Assessing the Impact of Hydration Status, Physical Activity, and Longevity on Nutromics Continuous Vancomycin Monitoring Device Performance

Adult participants are divided into seven cohorts. There will be up to 10 participants in each cohort. Each cohort is made up of a composition of different phases: • Vancomycin Exposure Phase (1000mg ~ 10 hour intravenous (IV) infusion) • Hydration Phase • Activity/Sweat Phase (Motion 1 – Arm Motion or Motion 2 – Bike Exercise) • Longevity Phase The following cohorts will be made up of the following phases: • Cohort 1 (Motion 1 + Hydration + Dose Day 2 (1000mg) + Longevity) • Cohort 2 (Dose Day 1 (1000mg) + Longevity) • Cohort 3 (Dose Day 1 (1000mg) + Motion 2 (Post-dose) + Dose Day 2 (1000mg) + Motion 1 (Post-dose) + Longevity) • Cohort 4 (Dosing (Day 1 – 3, 1000mg per day) + Longevity) • Cohort 5 (Hydration (Pre-Study) + Motion 1 Day 1 + Motion 2 Day 2) • Cohort 6 (Motion 1 Day 1 + Hydration + Longevity) • Cohort 7 (Hydration + Longevity) Dose/Dose day refers to a single 1000mg intravenous infusion of vancomycin. All cohorts will be required to attend an in-clinic visit no more than 5 consecutive days. Some cohorts may not require participants to attend all 5 in-clinic visits. Other cohorts may be contacted. Participants will be assigned to a cohort at screening. Participants may be enrolled into any cohort in a non-sequential manner; enrolment is not restricted to a predefined order. This flexible approach allows for parallel recruitment across cohorts based on eligibility and operational readiness. Participants may enrol up to 3 different cohorts within a 12 week period given that total volume of blood does not exceed 470mL from their participation in their first allocated cohort, up until their third cohort allocation. Participants must also be allocated to a different cohort after completion of their participation in a specific cohort. Participants should not repeat the same cohort throughout the entire study. This clinical study has a duration of up to 12 months. Screening Visit (~3 hours) During the Screening Visit, a member of the Study Team will provide the participant with the PICF for the participant to review. Participants will be enrolled in the study after the informed consent process is completed, provided they meet all inclusion criteria and none of the exclusion criteria. Each participant will be assigned a unique ID number, which will be documented on all study-related documents. The following procedures at the Screening Visit will be performed: • Demographic information (birth date, sex, race, and ethnicity) will be collected. • Review of medical history • Review of current medications, including herbal medicines, supplements, and vitamins; • Blood and Urine sample collection for relevant pathology tests (FBC, UEC, LFT and CRP), Serology, Urinalysis and Pregnancy test (if applicable) • Vital signs and Physical assessment for general health monitoring (including measurement of height and body weight) Participants in the Vancomycin dosing study arms must have a Creatinine Clearance (CrCl) greater than, or equal to 90mL/min. Serum creatinine will be tested at the Screening Visit, and the CrCl will be calculated using the Cockcroft-Gault formula. Once the eligibility of each participant is confirmed, they will be notified of their eligibility and booked in for a Dosing Visit. Participants that have enrolled and taken part in either dosing or non-dosing cohort, may re-screen for another cohort provided that their total blood volume collected does not exceed 470 mL over a 12-week period (equivalent to one standard blood donation). Re-screening blood collection may be waived, if the participant’s re-screening visit falls within 14 days of their previous screening visit, provided that their previous screening results remain within normal ranges and clinically acceptable. Baseline Procedures: Upon arrival at the Study Site for a clinic visit, participants will first undergo verification of eligibility and confirmation of informed consent. Baseline vital signs and relevant health information will be collected, and, if applicable (hydration phase), participants’ adherence to pre-visit instructions, including the food and fluid restriction after 10:00 PM, will be confirmed. Review of medical condition and any changes in medications. A blood or urine test will be taken to test for pregnancy in females of childbearing potential and relevant pathology tests are collected before any application of the Nutromics Sensor Devices. Up to 3 Nutromics Sensor Devices will be applied on each arm, a total of 6 devices per participant. An accelerometer sensor device will also be applied on to participants throughout the duration of the clinical study. The participants will complete a Pain Survey following the application of all the Nutromics Sensor Devices (+ 5 min). Assessment phase procedures will commence after 1 hour following application of the Nutromics Sensor Devices. Vancomycin Dosing (1000mg) Phase Cohorts that include Vancomycin phase within their study schedule will include the following procedures: • IV cannula (IVC) insertion into both arms - one for administering infusion, the other for blood collection • Blood PK collection at the following timepoints: o Prior to the administration of the vancomycin infusion (±15 mins), halfway (±15 mins and post-infusion (±15 mins) o 1 hr (±15 min) post-infusion timepoint o 3 hr (±15 min) post-infusion timepoint o 4 hr (±15 min) post-infusion timepoint o 6 hr (±30 min) post-infusion timepoint o 8 hr (±30 min) post-infusion timepoint (optional if additional phase procedures are performed on the same day) o 24 hr (± 30 min) post-infusion timepoint • Vital signs assessment performed during infusion Hydration Phase Participants in the Hydration Phase will undergo continuous device monitoring beginning in a dehydrated state, followed by a structured rehydration intervention. This allows direct comparison of vancomycin pharmacokinetics and sensor signal characteristics under both fluid conditions. Participants assigned to the Hydration Phase will only be allocated to either Day -1 (Pre-Study Visit) or Day 1 de-hydration procedures. • Participants enrolled in the Hydration Phase will be instructed to abstain from all oral fluid intake after 22:00 (10:00 PM) on the night prior to the following on-site study visit. • Participants will be reminded of this requirement during both the screening call and the Day prior to their clinic visit confirmation. • Compliance will be confirmed upon arrival via verbal confirmation and baseline vital signs. • Participants will receive a standardised volume of isotonic oral fluid (water and food if needed) to consume. Consumption can begin as soon as participants arrive on-site (depending on cohort allocation) the following day after cease of food and drink intake. o The full volume must be consumed within 30 minutes under direct supervision. • Continuous device monitoring will continue for a further 3 hours post-rehydration to assess hydration recovery dynamics and device signal quality. Activity/Sweat Phase (Motion 1 - Arm-motion, Motion 2 - Bike Exercise) The Activity and Sweat monitoring phase will assess the impact of physical activity and perspiration on Nutromics Sensor Device signal stability and response, and to characterise confounding signals related to sweat rate and composition. Participants will be allocated to either an arm-motion activity (motion 1) or a sweat motion activity (motion 2). Prior to either Activities: • Ensure devices are functioning and recording; log device IDs and start times. • Fit the participant with a Polar Verity Sense activity tracker (or equivalent heart rate monitoring device) on the arm. • Record baseline vital signs, skin temperature at sensor sites, and ambient room temperature/humidity. Standardised Activity Protocol – Arm Motion Activity (Motion 1) 1. Structured arm-motion activity (1.5 hours) — A video will be shown to participants for them to follow the standardised arm movements (repeated flexion/extension, abduction/adduction, simulated reaching) to create local mechanical perturbation at device sites. • Participants will be required to perform a selection or all the following motions: o Triceps Kickback o Bicep Curls o Reverse Flies o Triceps Overhead o Shoulder Extension o Shoulder internal rotation • Clinical site team may also perform the following motion on participants: o Device pressure test – using a blood pressure cuff to cover most if not all the devices on the upper arms at a consistent appropriate pressure o High frequency vibration test – using a massage gun on the participants 2-3cm away from the devices o Dislodgement test – Clinical team member will lightly pull (up, down, left and right) on applied devices on participants over clothing • 5 minutes per motion with 5 minutes of rest • A pain assessment will be requested to be completed upon each type of motion activity performed. Standardised Activity Protocol – Sweat Motion Activity (Motion 2) 1. Warm-up (5 minutes) — light cycle on an exercise bike. 2. Steady state - moderate cycle on an exercise bike (30 minutes) • Cycling at a speed to achieve and maintain ~60–70% age-predicted maximum heart rate (target HR = 0.6–0.7 × [220 - age] or a Rate of Perceived Exertion (RPE) 6-8). 3. Cool-down (5 minutes) — gradual reduction to resting HR. • Throughout the steady-state phase monitor HR (Polar Verity Sensor). • If HR is outside the target for >2 minutes, adjust workload to re-enter target zone. • Participant-reported RPE at 5-minute intervals during steady-state. • Collect a post-activity blood sample per study schedule. • Continue device monitoring through cooled-down period and until scheduled post-activity timepoint. • A subset or all the Nutromics Sensor devices and all other wearable materials will be removed. • Participants will complete a pain survey following removal of the Nutromics Sensor Devices (± 15mins). • Images are taken of the application sites for each of the Nutromics Sensor Device after their removal. • Up to 3 devices are applied per arm, a maximum of 6 devices per participant. Number of devices will vary which is dependent upon assessment phase arms. • Participants will complete a pain survey following application of all Nutromics Sensor Devices (± 15mins). Longevity Phase: During this phase, participants continue to wear the Nutromics Sensor Devices to evaluate signal stability, drift, and device performance over an extended wear period of up to 72 hours. Participants may have some devices removed over clinical trial period. For those that do not require devices to be removed until day 4 (or earlier), do not need to return to clinical site if their cohort schedule does not require them to return for device removal.,). For participants that do require device replacement throughout the clinical study, the following procedures will be performed for each study visit after the Day 1 of device application: • A subset of devices will be removed and replaced with new devices. The total number of devices after re-applying new devices will have no more than 3 devices per arm or no more than a total of 6. Once the subset of devices is removed, the following clinical procedures must be followed: • Pain assessment will be completed by the participant. • Images are taken of the application sites for each of the Nutromics Sensor Device following their removal. Care will be taken to avoid identifiable features. • The new subset of devices (same amount removed) will be re-applied onto the participant. • After re-application, the participant will complete a Pain Survey following the application of the new Nutromics Sensor Devices. Discharge Activities: Participants will be discharged from the study site at least 1 hour following the last study assessment phase and after a non-clinically significant vital sign assessment has been obtained. Pathology blood samples will be collected if not already performed. A vancomycin concentration testing blood sample will also be collected before discharge for participants in dosing cohorts (1 – 4). Participants may perform their normal daily activities including sports, showering (the accelerometer device is not water-proof and will need to be removed when showering), etc.; and the type of activity, start and end time of these activities shall be recorded by the participant. The Devices should not be submerged in water. Participants will be instructed to avoid apply excessive force or pressure on the Nutromics Sensor Devices. In the event the Nutromics Sensor Device(s) fall off, participants are instructed not re-apply the device and how to safely package the device for return to Nutromics. Return to clinic visits (subsequent clinic visits i.e. Day 2, 3 or 4): Participants will return to the site ~24 hours after Nutromics Sensor Device application. Clinical site team will discuss with participants on any impact from device application upon returning home, where the activity tracker/monitoring log (completed by the participant) is returned to the clinical site team. Participants allocated to the Hydration Phase for Day 2, their adherence to discharge instructions including the food and fluid restriction after 10:00 PM, will be confirmed. Before any removal of devices, baseline assessments including vital signs and pathology test bloods.